#METABOLOMICS WORKBENCH kaguilera_20210719_133918 DATATRACK_ID:2757 STUDY_ID:ST001902 ANALYSIS_ID:AN003094 PROJECT_ID:PR001197 VERSION 1 CREATED_ON August 11, 2021, 6:00 pm #PROJECT PR:PROJECT_TITLE Metabolomics analysis of AsPC-1 PDAC cells treated with Porcupine inhibitor PR:PROJECT_TITLE (LGK974) PR:PROJECT_SUMMARY WNT signaling promotes pancreatic ductal adenocarcinoma (PDAC) through diverse PR:PROJECT_SUMMARY effects on proliferation, differentiation, survival, and stemness. A subset of PR:PROJECT_SUMMARY PDAC with inactivating mutations in ring finger protein 43 (RNF43) have growth PR:PROJECT_SUMMARY dependency on autocrine WNT ligand signaling, which renders them susceptible to PR:PROJECT_SUMMARY porcupine inhibitors (PORCNi) that block WNT ligand acylation and secretion. For PR:PROJECT_SUMMARY this study, non-targeted metabolomic analyses were performed to explore the PR:PROJECT_SUMMARY therapeutic response of RNF43-mutant PDAC to the PORCNi LGK974. AsPC-1 PR:PROJECT_SUMMARY (RNF43-mutant) PDAC cells were treated with 25 nM LGK974 to explore stable PR:PROJECT_SUMMARY isotope-resolved metabolomics with uniform 1, D-glucose [U13-C6] labeling. PR:INSTITUTE UCLA PR:DEPARTMENT Pathology & Laboratory Medicine PR:LABORATORY Dawson Lab PR:LAST_NAME David PR:FIRST_NAME Dawson PR:ADDRESS 10833 Le Conte Avenue, Los Angeles, CA, 90095, USA PR:EMAIL ddawson@mednet.ucla.edu PR:PHONE (310) 267-2799 #STUDY ST:STUDY_TITLE Metabolomics analysis of AsPC-1 PDAC cells treated with Porcupine inhibitor ST:STUDY_TITLE (LGK974) ST:STUDY_SUMMARY WNT signaling promotes pancreatic ductal adenocarcinoma (PDAC) through diverse ST:STUDY_SUMMARY effects on proliferation, differentiation, survival, and stemness. A subset of ST:STUDY_SUMMARY PDAC with inactivating mutations in ring finger protein 43 (RNF43) have growth ST:STUDY_SUMMARY dependency on autocrine WNT ligand signaling, which renders them susceptible to ST:STUDY_SUMMARY porcupine inhibitors (PORCNi) that block WNT ligand acylation and secretion. For ST:STUDY_SUMMARY this study, non-targeted metabolomic analyses were performed to explore the ST:STUDY_SUMMARY therapeutic response of RNF43-mutant PDAC to the PORCNi LGK974. AsPC-1 ST:STUDY_SUMMARY (RNF43-mutant) PDAC cells were treated with 25 nM LGK974 to explore stable ST:STUDY_SUMMARY isotope-resolved metabolomics with uniform 1, D-glucose [U13-C6] labeling. ST:INSTITUTE University of California, Los Angeles ST:DEPARTMENT Pathology & Laboratory Medicine ST:LABORATORY Dawson Lab ST:LAST_NAME Dawson ST:FIRST_NAME David ST:ADDRESS 10833 LeConte Avenue ST:EMAIL ddawson@mednet.ucla.edu ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 6 ST:PHONE 310-825-0618 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - KA-010 Treatment:Control RAW_FILE_NAME=KA-010 SUBJECT_SAMPLE_FACTORS - KA-011 Treatment:Control RAW_FILE_NAME=KA-011 SUBJECT_SAMPLE_FACTORS - KA-012 Treatment:Control RAW_FILE_NAME=KA-012 SUBJECT_SAMPLE_FACTORS - KA-013 Treatment:25 nM LGK974 RAW_FILE_NAME=KA-013 SUBJECT_SAMPLE_FACTORS - KA-014 Treatment:25 nM LGK974 RAW_FILE_NAME=KA-014 SUBJECT_SAMPLE_FACTORS - KA-016 Treatment:25 nM LGK974 RAW_FILE_NAME=KA-016 #COLLECTION CO:COLLECTION_SUMMARY AsPC-1 PDAC cells were treated with DMSO vehicle control or 25 nM LGK974 (in CO:COLLECTION_SUMMARY triplicate per group). 1 x 10^6 cells were washed with ice-cold 150 mM ammonium CO:COLLECTION_SUMMARY acetate twice before adding 1 mL of ice-cold 80% methanol. After vigorous CO:COLLECTION_SUMMARY vortexing, samples were centrifuged at maximum speed, the aqueous layer was CO:COLLECTION_SUMMARY transferred to a glass vial and the metabolites were dried under vacuum. CO:COLLECTION_SUMMARY Metabolites were resuspended in 50 μL 70% acetonitrile (ACN) and 5 μL of this CO:COLLECTION_SUMMARY solution used for the mass spectrometer-based analysis. The analysis was CO:COLLECTION_SUMMARY performed on a Q Exactive (Thermo Fisher Scientific) in polarity-switching mode CO:COLLECTION_SUMMARY with positive voltage 4.0 kV and negative voltage 4.0 kV. The mass spectrometer CO:COLLECTION_SUMMARY was coupled to an UltiMate 3000RSLC (Thermo Fisher Scientific) UHPLC system. CO:COLLECTION_SUMMARY Mobile phase A was 5 mM ammonium acetate (NH4AcO), pH 9.9, B was acetonitrile CO:COLLECTION_SUMMARY and the separation achieved on a Luna 3 mm NH2 100 A column (150 × 2.0 mm, CO:COLLECTION_SUMMARY Phenomenex). The flow was 200 μL/min, and the gradient ran from 15% A to 95% A CO:COLLECTION_SUMMARY in 18 min, followed by an isocratic step for 9 min and re-equilibration for 7 CO:COLLECTION_SUMMARY min. Metabolites were detected and quantified as area under the curve based on CO:COLLECTION_SUMMARY retention time and accurate mass (≤ 3 p.p.m.) using the TraceFinder 3.1 CO:COLLECTION_SUMMARY (Thermo Fisher Scientific). Relative amounts of metabolites between various CO:COLLECTION_SUMMARY conditions, as well as percentage of [13C6] glucose labelling, were calculated CO:COLLECTION_SUMMARY and corrected for naturally occurring 13C abundance. CO:SAMPLE_TYPE Cultured cells #TREATMENT TR:TREATMENT_SUMMARY AsPC-1 PDAC cells were plated at 500,000 cells per well in 6 well plates in TR:TREATMENT_SUMMARY ultra-low attachment conditions. Cells were pre-treated with 25 nM LGK974 or TR:TREATMENT_SUMMARY DMSO vehicle control for 12 hours, maintained in normal media conditions. Media TR:TREATMENT_SUMMARY was exchanged with [13C6] glucose labeling media and cells were re-treated with TR:TREATMENT_SUMMARY 25 nM LGK974 or DMSO vehicle control for 24 hours. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Cells were washed with ice-cold 150 mM ammonium acetate twice before adding 1 mL SP:SAMPLEPREP_SUMMARY of ice-cold 80% methanol. After vigorous vortexing, samples were centrifuged at SP:SAMPLEPREP_SUMMARY maximum speed, the aqueous layer was transferred to a glass vial and the SP:SAMPLEPREP_SUMMARY metabolites were dried under vacuum. Metabolites were resuspended in 50 μL 70% SP:SAMPLEPREP_SUMMARY acetonitrile (ACN) and 5 μL of this solution used for the mass SP:SAMPLEPREP_SUMMARY spectrometer-based analysis. The analysis was performed on a Q Exactive (Thermo SP:SAMPLEPREP_SUMMARY Fisher Scientific) in polarity-switching mode with positive voltage 4.0 kV and SP:SAMPLEPREP_SUMMARY negative voltage 4.0 kV. The mass spectrometer was coupled to an UltiMate SP:SAMPLEPREP_SUMMARY 3000RSLC (Thermo Fisher Scientific) UHPLC system. Mobile phase A was 5 mM SP:SAMPLEPREP_SUMMARY ammonium acetate (NH4AcO), pH 9.9, B was acetonitrile and the separation SP:SAMPLEPREP_SUMMARY achieved on a Luna 3 mm NH2 100 A column (150 × 2.0 mm, Phenomenex). The flow SP:SAMPLEPREP_SUMMARY was 200 μL/min, and the gradient ran from 15% A to 95% A in 18 min, followed by SP:SAMPLEPREP_SUMMARY an isocratic step for 9 min and re-equilibration for 7 min. Metabolites were SP:SAMPLEPREP_SUMMARY detected and quantified as area under the curve based on retention time and SP:SAMPLEPREP_SUMMARY accurate mass (≤ 3 p.p.m.) using the TraceFinder 3.1 (Thermo Fisher SP:SAMPLEPREP_SUMMARY Scientific). Relative amounts of metabolites between various conditions, as well SP:SAMPLEPREP_SUMMARY as percentage of [13C6]glucose labeling, were calculated and corrected for SP:SAMPLEPREP_SUMMARY naturally occurring 13C abundance. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Mobile phase A was 5 mM ammonium acetate (NH4AcO), pH 9.9, B was acetonitrile CH:CHROMATOGRAPHY_SUMMARY and the separation achieved on a Luna 3 mm NH2 100 A column (150 × 2.0 mm, CH:CHROMATOGRAPHY_SUMMARY Phenomenex). The flow was 200 μL/min, and the gradient ran from 15% A to 95% A CH:CHROMATOGRAPHY_SUMMARY in 18 min, followed by an isocratic step for 9 min and re-equilibration for 7 CH:CHROMATOGRAPHY_SUMMARY min. CH:CHROMATOGRAPHY_TYPE Normal phase CH:INSTRUMENT_NAME Q Exactive CH:COLUMN_NAME Luna 3 mm NH2 100 A CH:FLOW_RATE 200 uL/min CH:SOLVENT_A 5 mM ammonium acetate (NH4AcO), pH 9.9 CH:SOLVENT_B acetonitrile #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE API MS:ION_MODE NEGATIVE MS:MS_COMMENTS Metabolites were detected and quantified as area under the curve based on MS:MS_COMMENTS retention time and accurate mass (≤ 3 p.p.m.) using the TraceFinder 3.1 MS:MS_COMMENTS (Thermo Fisher Scientific). Relative amounts of metabolites between various MS:MS_COMMENTS conditions, as well as percentage of [13C6] glucose labeling, were calculated MS:MS_COMMENTS and corrected for naturally occurring 13C abundance. MS:ION_SPRAY_VOLTAGE 4000 #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS area MS_METABOLITE_DATA_START Samples KA-010 KA-011 KA-012 KA-013 KA-014 KA-016 Factors Treatment:Control Treatment:Control Treatment:Control Treatment:25 nM LGK974 Treatment:25 nM LGK974 Treatment:25 nM LGK974 2-Oxoadipate M0 1013239 252001 343304 251417 NA 499638 5-Oxoproline M0 35813218 42272952 39952544 33325723 22898313 21104363 5-Oxoproline M1 1803893 1974167 1980506 1493688 862658 336914 5-Oxoproline M2 1866482 1395782 1060355 1298829 764480 688638 5-Oxoproline M3 294302 192962 234472 321295 83587 NA 5-Oxoproline M4 117182 72522 217199 NA NA NA 5-Oxoproline M5 244063 173171 111870 NA NA NA Homocysteine M0 NA NA NA 2291527 2809009 3229201 Norvaline 106612254 108732552 101727866 68637578 67188297 70908844 Orotidine M5 212225 413212 1110411 NA NA NA Orotidine M8 NA NA 106023 NA NA NA ATP M7 NA 85074 NA NA NA NA NADP+ 582637 490629 1104297 264219 680522 188945 dCMP M0 793974 454126 372476 947525 726851 915723 dCMP M1 154204 166168 51802 263080 243360 47638 dCMP M2 2038225 1249631 1724134 2120205 1939154 2611274 dTMP M2 12724152 7395052 9832887 11541077 10262635 18478785 dTMP M3 63296 NA 129169 NA NA 181653 dTMP M4 NA NA 110680 NA NA NA dU M0 NA NA NA NA 134735 NA dU M4 194506 127931 NA NA NA NA Phosphoethanolamine M0 4432640 4475447 4643949 2682846 2246817 2286274 Hexose phosphate M0 2318450 2481249 2992976 2134117 2137913 1521677 Hexose phosphate M5 NA NA NA NA NA 70319 Hexose phosphate M6 818035 721043 926729 581227 588007 564927 Inositol 1503302723 1495585612 1453787387 986602273 895969554 910389243 Pyruvate M0 NA NA NA 79409 76189 NA Serine Std 142419433 133969796 119602419 131065294 142559986 62083073 Sorbitol M0 NA 9297907 8378440 NA NA NA Sorbitol M6 807884 2238103 2456074 NA NA 126070 UDP-Glucose M0 524770 301813 442235 375541 777351 391369 UDP-Glucose M10 1113559 634722 841600 104701 169155 123082 UDP-Glucose M11 4613075 4965754 5466200 1565708 1438127 1571333 UDP-Glucose M12 1157270 1035744 1374769 NA NA 70062 UDP-Glucose M13 873332 1485594 1178068 91102 214063 67022 UDP-Glucose M14 176198 383185 491532 NA NA NA UDP-Glucose M3 350668 77432 NA 84746 171465 NA UDP-Glucose M4 NA 80068 85689 NA 108800 NA UDP-Glucose M5 1198595 1546268 1701914 257552 678668 289418 UDP-Glucose M6 3080695 3537438 3018776 1924987 1748000 1531688 UDP-Glucose M7 710404 688469 663500 230666 327835 79963 UDP-Glucose M8 597401 552615 326268 93072 153856 NA UDP-Glucose M9 NA 70345 NA NA NA NA GlcNAc-6-phosphate M7 76529 NA 80647 122702 90607 NA GlcNAc-6-phosphate M8 628390 921549 431028 NA 81703 77125 Glucuronic Acid M3 91631 NA NA NA NA NA Glucuronic Acid M4 74729 70822 200182 279283 NA 151096 Glucuronic Acid M5 84227 92793 156602 NA NA NA Glucuronic Acid M6 2884019 3497952 2892652 1059345 1766164 1232383 UDP-GlcNAc M10 NA 279774 234466 NA NA NA UDP-GlcNAc M11 2934459 2719668 2115139 539302 712247 785383 UDP-GlcNAc M12 681825 1015840 1014420 NA 208194 158242 UDP-GlcNAc M13 2765025 3015732 3361682 660594 341643 342594 UDP-GlcNAc M14 930163 886785 607979 NA NA NA UDP-GlcNAc M15 405584 567248 660360 NA NA NA UDP-GlcNAc M7 655896 738290 487947 253174 150338 NA UDP-GlcNAc M8 1442570 2170909 2154069 590138 621213 197251 UDP-GlcNAc M9 82829 NA 275700 71183 NA 72597 CDP M0 493941 284707 131114 184762 NA 453760 CDP M5 75048 242335 96370 NA NA NA CMP M0 102306 69686 116617 95967 NA 137443 CTP M0 608816 382166 466955 151858 353079 197133 CTP M5 596211 374785 868617 NA NA NA GDP M0 1153586 488848 791746 1462074 1125890 1298388 GDP M4 104340 NA 51108 NA NA NA GDP M5 2294951 1462377 1979439 813699 308539 405681 GMP M0 320851 NA 167567 78588 299348 461912 GMP M5 586015 165065 647132 75438 NA 180173 GTP M0 1402821 600240 1605019 859495 754975 194007 GTP M5 3254720 2504664 3735877 296177 422072 327793 T M0 NA NA NA 226981 109921 91903 Thymine M0 1028096 1706908 1836759 2008919 NA 739387 UDP M0 3568784 3109206 3289819 3111821 2616782 2974806 UDP M1 137460 63578 NA 86391 107965 NA UDP M2 NA 188130 64386 57937 NA 182133 UDP M4 336969 502962 110615 147690 NA 76947 UDP M5 7103914 5616591 5245352 2249850 1532460 2994115 UDP M6 1313948 962022 846893 205255 90494 218229 UDP M7 1336640 1289474 1479007 158210 72854 309463 UDP M8 703719 339729 881109 NA NA NA UTP M0 4319701 3986244 3908425 2586326 2564078 1888015 UTP M1 68920 303158 144948 128794 81109 NA UTP M2 130073 211370 148777 NA NA NA UTP M4 268309 192027 235909 NA NA NA UTP M5 8045793 6980807 7824869 1890282 2446852 1676692 UTP M6 1281807 1657697 1866970 67942 NA 64851 UTP M7 1903523 1754742 2230728 86161 235688 168800 UTP M8 441513 408847 467276 NA NA NA IMP M0 287117 293236 467413 451074 195018 524831 IMP M5 1488019 1093488 1998824 474596 624215 558038 PRPP M0 182749 61891 353896 NA NA NA PRPP M5 2965837 723347 2797790 298174 81321 65283 UMP M0 610188 158336 533618 366944 81510 629769 UMP M5 1484196 663743 578013 92787 NA 288485 UMP M6 61545 NA 156975 NA NA NA UMP M7 103061 NA 98133 NA NA NA Uracil M0 24835978 25167713 28190424 23863162 NA 26235181 Uracil M1 3256855 3173417 3916929 1665061 2205327 2342171 Uracil M2 3631757 3547352 4247707 650482 337908 1143405 Uracil M3 1403949 1488593 2323616 504089 NA 325268 Xanthine M0 2753219 4250025 5241275 2890744 2328906 3480177 2-HG M0 8527049 11015166 9355027 5715675 6037689 5864289 2-HG M1 NA 142223 179641 NA NA NA 2-HG M2 185041 NA 303784 NA NA NA Aconitic acid M0 992605 1237374 1650419 254560 243797 395443 Aconitic acid M1 151763 251674 272100 NA NA NA Aconitic acid M2 278128 260360 485248 NA NA NA Aconitic acid M3 NA 114005 187377 NA NA NA Glutamate Std 106556713 109180879 104787981 96972751 115568444 85956452 Pantothenic acid (Vitamin B5) 30471503 32746515 33908284 20036752 17983242 17026351 Phosphate 934295805 622696115 718504686 938933131 679828372 1444567103 Pyrophosphate 8076785 2920227 6012526 2252637 1458332 3321852 Thiosulfate 358852 896105 500006 499840 327630 293713 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID Formula m/z RT Polarity 2-Oxoadipate M0 C00322 C6H8O5 159.0299 16.2 negative 5-Oxoproline M0 C01879 C5H7NO3 128.0353 9.8 negative 5-Oxoproline M1 C01879 C5H7NO3 129.0387 9.8 negative 5-Oxoproline M2 C01879 C5H7NO3 130.042 9.8 negative 5-Oxoproline M3 C01879 C5H7NO3 131.0454 9.8 negative 5-Oxoproline M4 C01879 C5H7NO3 132.0487 9.8 negative 5-Oxoproline M5 C01879 C5H7NO3 133.0521 9.8 negative Homocysteine M0 C00155 C4H9NO2S 135.0315 7.9 negative Norvaline C01799 C5H11NO2 116.0717 7.1 negative Orotidine M5 X00006 C10H12N2O8 292.0688 13 negative Orotidine M8 X00006 C10H12N2O8 295.0789 13 negative ATP M7 C00002 C10H16N5O13P3 513.0119 23 negative NADP+ C00006 C21H28N7O17P3 742.0682 18.3 negative dCMP M0 C00239 C9H14N3O7P 306.0497 17 negative dCMP M1 C00239 C9H14N3O7P 307.053 17 negative dCMP M2 C00239 C9H14N3O7P 308.0564 17 negative dTMP M2 C00364 C10H14N2O8 323.056 16.5 negative dTMP M3 C00364 C10H14N2O9 324.0594 16.5 negative dTMP M4 C00364 C10H14N2O10 325.0627 16.5 negative dU M0 C00526 C9H12N2O5 227.0673 3.9 negative dU M4 C00526 C9H12N2O5 231.0807 3.9 negative Phosphoethanolamine M0 C00346 C2H8NO4P 140.0118 14 negative Hexose phosphate M0 C00085 C6H13O9P 259.0224 16 negative Hexose phosphate M5 C00085 C6H13O9P 264.0392 16 negative Hexose phosphate M6 C00085 C6H13O9P 265.0425 16 negative Inositol C00137 C6H12O6 179.0561 6.4 negative Pyruvate M0 C00022 C3H4O3 87.0088 7 negative Serine Std C00065 C3H7NO3 108.0424 9.3 negative Sorbitol M0 C00794 C6H14O6 181.0718 5.5 negative Sorbitol M6 C00794 C6H14O6 187.0919 5.5 negative UDP-Glucose M0 C00029 C15H24N2O17P2 565.0477 16.6 negative UDP-Glucose M10 C00029 C15H24N2O17P12 575.0812 16.6 negative UDP-Glucose M11 C00029 C15H24N2O17P13 576.0846 16.6 negative UDP-Glucose M12 C00029 C15H24N2O17P14 577.0879 16.6 negative UDP-Glucose M13 C00029 C15H24N2O17P15 578.0913 16.6 negative UDP-Glucose M14 C00029 C15H24N2O17P16 579.0946 16.6 negative UDP-Glucose M3 C00029 C15H24N2O17P5 568.0578 16.6 negative UDP-Glucose M4 C00029 C15H24N2O17P6 569.0611 16.6 negative UDP-Glucose M5 C00029 C15H24N2O17P7 570.0645 16.6 negative UDP-Glucose M6 C00029 C15H24N2O17P8 571.0678 16.6 negative UDP-Glucose M7 C00029 C15H24N2O17P9 572.0712 16.6 negative UDP-Glucose M8 C00029 C15H24N2O17P10 573.0745 16.6 negative UDP-Glucose M9 C00029 C15H24N2O17P11 574.0779 16.6 negative GlcNAc-6-phosphate M7 C00357 C8H16NO9P 307.0724 16.2 negative GlcNAc-6-phosphate M8 C00357 C8H16NO9P 308.0758 15.5 negative Glucuronic Acid M3 C00191 C6H10O7 196.0454 13 negative Glucuronic Acid M4 C00191 C6H10O7 197.0488 13 negative Glucuronic Acid M5 C00191 C6H10O7 198.0521 13 negative Glucuronic Acid M6 C00191 C6H10O7 199.0555 13 negative UDP-GlcNAc M10 C00043 C17H27N3O17P2 616.1078 16.3 negative UDP-GlcNAc M11 C00043 C17H27N3O17P2 617.1111 16.3 negative UDP-GlcNAc M12 C00043 C17H27N3O17P2 618.1145 16.3 negative UDP-GlcNAc M13 C00043 C17H27N3O17P2 619.1178 16.3 negative UDP-GlcNAc M14 C00043 C17H27N3O17P2 620.1212 16.3 negative UDP-GlcNAc M15 C00043 C17H27N3O17P2 621.1245 16.3 negative UDP-GlcNAc M7 C00043 C17H27N3O17P2 613.0977 16.3 negative UDP-GlcNAc M8 C00043 C17H27N3O17P2 614.1011 16.3 negative UDP-GlcNAc M9 C00043 C17H27N3O17P2 615.1044 16.3 negative CDP M0 C00112 C9H15N3O11P2 402.0109 19.1 negative CDP M5 C00112 C9H15N3O11P2 407.0277 19.1 negative CMP M0 C00055 C9H14N3O8P 322.0446 16.3 negative CTP M0 C00063 C9H16N3O14P3 481.9772 21.5 negative CTP M5 C00063 C9H16N3O14P3 486.994 21.5 negative GDP M0 C00035 C10H15N5O11P2 442.0171 21 negative GDP M4 C00035 C10H15N5O11P2 446.0305 21 negative GDP M5 C00035 C10H15N5O11P2 447.0338 21 negative GMP M0 C00144 C10H14N5O8P 362.0507 17.5 negative GMP M5 C00144 C10H14N5O8P 367.0675 17.5 negative GTP M0 C00044 C10H16N5O14P3 521.9834 25 negative GTP M5 C00044 C10H16N5O14P3 527.0001 25 negative T M0 C00214 C10H14N2O5 241.083 3.5 negative Thymine M0 C00178 C5H6N2O2 125.0357 3.2 negative UDP M0 C00015 C9H14N2O12P2 402.9949 19.5 negative UDP M1 C00015 C9H14N2O12P2 403.9983 19.5 negative UDP M2 C00015 C9H14N2O12P2 405.0016 19.5 negative UDP M4 C00015 C9H14N2O12P2 407.0083 19.5 negative UDP M5 C00015 C9H14N2O12P2 408.0117 19.5 negative UDP M6 C00015 C9H14N2O12P2 409.015 19.5 negative UDP M7 C00015 C9H14N2O12P2 410.0184 19.5 negative UDP M8 C00015 C9H14N2O12P2 411.0217 19.5 negative UTP M0 C00075 C9H15N2O15P3 482.9613 22.5 negative UTP M1 C00075 C9H15N2O15P3 483.9646 22.5 negative UTP M2 C00075 C9H15N2O15P3 484.968 22.5 negative UTP M4 C00075 C9H15N2O15P3 486.9747 22.5 negative UTP M5 C00075 C9H15N2O15P3 487.978 22.5 negative UTP M6 C00075 C9H15N2O15P3 488.9814 22.5 negative UTP M7 C00075 C9H15N2O15P3 489.9847 22.5 negative UTP M8 C00075 C9H15N2O15P3 490.9881 22.5 negative IMP M0 C00130 C10H13N4O8P 347.0398 18 negative IMP M5 C00130 C10H13N4O8P 352.0566 18 negative PRPP M0 C00119 C5H13O14P3 388.9445 24 negative PRPP M5 C00119 C5H13O14P3 393.9613 24 negative UMP M0 C00105 C9H13N2O9P 323.0286 16.5 negative UMP M5 C00105 C9H13N2O9P 328.0453 16.5 negative UMP M6 C00105 C9H13N2O9P 329.0487 16.5 negative UMP M7 C00105 C9H13N2O9P 330.052 16.5 negative Uracil M0 C00106 C4H4N2O2 111.02 3.75 negative Uracil M1 C00106 C4H4N2O2 112.0234 3.75 negative Uracil M2 C00106 C4H4N2O2 113.0267 3.75 negative Uracil M3 C00106 C4H4N2O2 114.0301 3.75 negative Xanthine M0 C00385 C5H4N4O2 151.0262 11.5 negative 2-HG M0 C02630 C5H8O5 147.0299 15.5 negative 2-HG M1 C02630 C5H8O5 148.0333 15.5 negative 2-HG M2 C02630 C5H8O5 149.0366 15.5 negative Aconitic acid M0 C00417 C6H6O6 173.0092 19 negative Aconitic acid M1 C00417 C6H6O6 174.0125 19 negative Aconitic acid M2 C00417 C6H6O6 175.0159 19 negative Aconitic acid M3 C00417 C6H6O6 176.0192 19 negative Glutamate Std C00025 C5H9NO5 152.0597 13.5 negative Pantothenic acid (Vitamin B5) C00864 C9H17NO5 218.1034 10.2 negative Phosphate C00009 H3PO4 96.9696 16.8 negative Pyrophosphate C00013 H4O7P2 176.936 23 negative Thiosulfate C00320 H2S2O3 112.9373 17 negative METABOLITES_END #END